Gene mutation site group of NK/T cell lymphoma, targeted sequencing kit and application

A technology for targeted sequencing and lymphoma. It is applied in recombinant DNA technology, DNA/RNA fragments, and the determination/inspection of microorganisms. The effect of competence and efficiency

Pending Publication Date: 2020-10-30
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Comparative genomic hybridization detection showed that NK / TCL had deletions on chromosomes such as 6q, which resulted in the silencing of tumor suppressor genes such as PRDM1, ATG5, AIM1, FOXO3 and HACE

Method used

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  • Gene mutation site group of NK/T cell lymphoma, targeted sequencing kit and application
  • Gene mutation site group of NK/T cell lymphoma, targeted sequencing kit and application
  • Gene mutation site group of NK/T cell lymphoma, targeted sequencing kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The reagents used in this example are specifically shown in Table 3, and other reagents are commercially available reagents unless otherwise specified.

[0063] table 3

[0064]

[0065]

[0066] 1. Sample acquisition

[0067] 102 patients with NK / T cell lymphoma were willing to undergo molecular testing and obtained informed consent. Frozen tissue samples, paraffin tissue samples, or biopsy tissue samples were obtained from the patients.

[0068] 2. Sample DNA extraction process:

[0069] ① DNA extraction reagents for frozen specimens were purchased from Promega (Beijing, China), and the specific steps are as follows:

[0070] Put the tumor tissue into a 1.5mL centrifuge tube containing 600 μL of nuclear lysate and 20 μL of proteinase K, and incubate in a metal bath at 56°C until the tumor tissue is completely digested (no longer than 8 hours). Add 3 μL RNase to the above sample, and incubate in a 36°C metal bath for 15-30 minutes. Add 200 μL of protein sedim...

Embodiment 2

[0151] Example 2 result verification

[0152] A high success rate PCR enzyme KOD FX kit (TOYOBO, Osaka, Japan) was used to perform a PCR experiment to sequence the mutation site of one of the DNA samples of the NK / T cell lymphoma patient in Example 1 to verify the experimental results.

[0153] Specifically, the reaction system of PCR is as shown in Table 12:

[0154] Table 12

[0155] Reagent Dosage Final concentration 2x PCR buffer for KOD FX 25 μL 1× 2mM dNTPs 10μL 0.4mM each 10pmol / μL Primer#1 1.5μL 0.3μM 10pmol / μL Primer#2 1.5μL 0.3μM template DNA 10μL KOD FX (1.0U / μL) 1μL 1.0U / 50μL Autoclaved, distilled water Up to 50μL

[0156] The reaction conditions of PCR are as shown in table 13:

[0157] Table 13

[0158]

[0159] The obtained sequencing results are as figure 1 shown. Will figure 1 The sequencing results shown are consistent with the human genome sequence hg19 in the ucsc databas...

Embodiment 3

[0161] Embodiment 3 Judging prognosis and guiding medication

[0162] 1. Patients with MGA mutations have higher disease stages and poorer prognosis.

[0163] 2. Patients with mutations in genes related to epigenetic acetylation regulation such as EP300, HDAC9, ARID1A are sensitive to targeted drug Chidmide treatment.

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a gene mutation site group of NK/T cell lymphoma and a kit and application thereof. The invention provides a gene mutation site group of NK/T cell lymphoma, which can be used for evaluating molecular typing of NK/T cell lymphoma, predicting prognosis of a patient and guiding medication of the patient. According tothe kit, an IlluminaMiseq sequencing platform and performance sequencing are adopted, a series of mutant genes related to NK/TCL diseases can be efficiently and accurately detected, the detection period is short, the detection cost is low, the application range is wide, and the accuracy is high.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a group of gene mutation site groups of NK / T cell lymphoma, a kit and application thereof. Background technique [0002] NK / T-cell lymphoma (NK / TCL) is a kind of CD56-positive / cytoplasmic CD3-positive lymphocyte malignant proliferation disease, which occurs in Asia and is closely related to Epstein-Barr virus (EB virus) infection. NK / TCL is highly malignant, highly invasive, resistant to conventional chemotherapy, and has a very poor prognosis. The five-year survival rate of advanced patients is only 20%. heavy burden and pain. [0003] The malignant degree of NK / TCL is high, the prognosis of patients is poor, and the pathogenesis has yet to be elucidated. Previous cytogenetic studies have shown that NK / TCL exhibits significant genomic and epigenetic instability. Comparative genomic hybridization detection showed that NK / TCL had deletions on chromosomes such as 6q, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6869C12N15/11
CPCC12Q1/6886C12Q1/6869C12Q2600/156
Inventor 赵维莅熊杰王黎刘以哲俞浩
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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